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Method for preparing decellularized biological tissue material

A biological tissue and decellularization technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve the problems of residual toxic substances, tissue damage, incomplete cell removal, etc., to reduce damage, low equipment requirements, and reduce Effect of risk of tissue calcification

Inactive Publication Date: 2020-05-08
SHANGHAI MICROPORT CARDIOFLOW MEDTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention provides a method for preparing decellularized biological tissue materials, which is used to solve the problems of incomplete cell removal, serious tissue damage or residual toxic substances in the preparation of decellularized biological tissue materials by the current method

Method used

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  • Method for preparing decellularized biological tissue material
  • Method for preparing decellularized biological tissue material
  • Method for preparing decellularized biological tissue material

Examples

Experimental program
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Effect test

Embodiment 1

[0051] Step 1: Spread a layer of sterile non-woven fabric evenly in a clean sterilized square pan, add sterile phosphate buffer to soak the non-woven fabric, and spread the pretreated bovine pericardium tissue on the soaked non-woven fabric Gently peel away the obvious fatty tissue layer without damaging the fibrous layer. Then, with the rough surface (fibrous layer) of the biological tissue facing up, gently wipe with a dust-free cloth to remove the fat on the surface of the tissue. Cut off the irregular parts and free fibers of the biological tissue, cut into small pieces (15mmx15mm), wash with normal saline at room temperature for 3 times, each time for 3 to 5 minutes;

[0052] Step 2, freeze-thaw treatment: the biological tissue treated in step 1 was pre-cooled at 4°C for 10 minutes, frozen at -80°C for 2 hours, rewarmed in a 37°C water bath for 15 minutes, and this process was repeated 3 times;

[0053] Step 3, ultra-high pressure treatment: thoroughly rinse the biologic...

Embodiment 2

[0069] Step 1: Spread a layer of sterile non-woven fabric evenly in a clean sterilized square pan, add sterile phosphate buffer saline to soak the non-woven fabric, and spread the pretreated porcine pericardium tissue on the soaked non-woven fabric Gently peel away the obvious fatty tissue layer without damaging the fibrous layer. Then, with the rough surface (fibrous layer) of the biological tissue facing up, gently wipe with a dust-free cloth to remove the fat on the surface of the tissue. Cut off the irregular parts and free fibers of the biological tissue, cut into small pieces (15mmx15mm), wash with normal saline at room temperature for 3 times, each time for 3 to 5 minutes;

[0070] Step 2, freeze-thaw treatment: the biological tissue treated in step 1 was pre-cooled at 4°C for 15 minutes, frozen at -80°C for 1.5 hours, rewarmed in a 37°C water bath for 15 minutes, and this process was repeated 3 times;

[0071] Step 3, ultra-high pressure treatment: thoroughly rinse th...

Embodiment 3

[0088] Step 1: Spread a layer of sterile non-woven fabric evenly in a clean sterilized square pan, add sterile phosphate buffer saline to soak the non-woven fabric, and spread the pretreated porcine pericardium tissue on the soaked non-woven fabric Gently peel away the obvious fatty tissue layer without damaging the fibrous layer. Then, with the rough surface (fibrous layer) of the biological tissue facing up, gently wipe with a dust-free cloth to remove the fat on the surface of the tissue. Cut off the irregular parts and free fibers of the biological tissue, cut into small pieces (15mmx15mm), wash with normal saline at room temperature for 3 times, each time for 3 to 5 minutes;

[0089] Step 2, freeze-thaw treatment: the biological tissue treated in step 1 was pre-cooled at 4°C for 20 minutes, frozen at -80°C for 2 hours, rewarmed in a 37°C water bath for 15 minutes, and this process was repeated 3 times;

[0090] Step 3, ultra-high pressure treatment: thoroughly rinse the bi...

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Abstract

The invention belongs to the technical field of biological tissue materials, and particularly relates to a method for preparing a decellularized biological tissue material. The method comprises the following steps: three processes including freezing and thawing treatment, ultrahigh pressure treatment and nucleic acid removal, wherein the ultrahigh pressure treatment is carried out under a pressureof 400-600 MPa. Compared with the prior art, a repeated freezing and thawing and ultrahigh pressure combined method is adopted in the technical scheme, the time consumed in the process can be remarkably shortened, the decellularization efficiency can be improved, in addition, experimental parameters are milder, and the damage to a biological scaffold structure is reduced. The pressure adopted inthe ultrahigh pressure treatment in the embodiment cannot cause a critical ice phenomenon of medium water, and the integrity and mechanical properties of a tissue structure can be well reserved.

Description

technical field [0001] The invention belongs to the technical field of biological tissue materials, and in particular relates to a method for preparing decellularized biological tissue materials. Background technique [0002] Due to their low immunogenicity, intact fibrous structure, and good mechanical strength, decellularized biological tissue materials have received increasing attention and applications in the field of medical devices. [0003] At present, the commonly used decellularization methods mainly include physical methods, enzyme digestion methods, acid-base methods, and chemical detergent methods. However, a large number of studies have shown that there are many problems in the above methods: (1) The physical method has poor decellularization effect, which is easy to cause cells and DNA residues, causing the body's immune response; (2) Enzyme digestion method requires a higher concentration of enzyme , high concentration of enzymes can easily cause damage to co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
CPCA61L27/3604A61L27/3687A61L27/3691A61L2430/40
Inventor 何海红尹宗明赵婧桂宝珠陈国明李雨
Owner SHANGHAI MICROPORT CARDIOFLOW MEDTECH CO LTD